Anti-inflammatory peptides derived from IL-2 and analogues thereof

Abstract

Synthetic anti-inflammatory peptides derived from the sequence of IL-2 are provided. Parent peptides of the sequences Ile-Val-Leu, Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:1) and Arg-Met-Leu-Thr (SEQ ID NO:2), were obtained by elastase enzymatic digestion of IL-2, synthesized and modified. The peptides are useful in conditions of acute and chronic inflammation such as in autoimmune diseases.

1. A synthetic anti-inflammatory peptide of IL-2 or an anti-inflammatory derivative thereof, which inhibits in vitro: (i) adhesion of activated T cells to fibronectin, laminin and/or collagen-type IV; (ii) chemotactic migration of T cells through fibronectin; and/or (iii) spontaneous or TNF-α-induced secretion of IL-8 or IL-1β, from intestinal epithelial cells, selected from the group consisting of:
 (i) peptides pep1, pep2, and pep3 consisting of the sequences: 
 (pep1) Ile-Val-Leu  
 (pep2) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:1)  
 (pep3) Arg-Met-Leu-Thr (SEQ ID NO:2)  
 
 (ii) peptides obtained from pep2 by deletion of one or more amino acid residues;  
 (iii) peptides obtained by addition to peptides (i) or (ii) of one or more natural or non-natural amino acid residues to the N-terminus and/or C-terminus;  
 (iv) peptides obtained by replacement of one amino acid residue of peptides (i) to (iii) by another natural amino acid residue or by a non-natural amino acid residue;  
 (v) peptides of (i) to (iii) which are all-L, all-D or a combination of D- and L-amino acid residues;  
 (vi) amide derivatives of the C-terminal residue of peptides (i) to (v);  
 (vii) cyclic derivatives of peptides (i) to (vi) in which the peptide is cyclized by an intramolecular bond; and  
 (viii) dual peptides consisting of two of the same or combination of peptides (i) to (vii), wherein the two peptides are covalently linked to one another directly or through a spacer.  
 
     
     
       2. The synthetic peptide Ile-Val-Leu (pep1) and derivatives thereof according to  claim 1 , obtained by:
 (a) elongation by up to 3-4 further amino acid residues at the N- and/or C-terminal;  
 (b) substitution of the Ile residue by a natural or non-natural amino acid hydrophilic polar neutral or negatively charged, or hydrophobic non-polar neutral amino acid residue;  
 (c) substitution of the Val residue by a hydrophobic, non-charged natural or non-natural amino acid residue;  
 (d) substitution of the Leu residue by a hydrophobic, non-charged natural or non-natural amino acid residue;  
 (e) amidation of the C-terminal Leu residue;  
 (f) cyclization of pep1 or of any peptide of (a) to (e); or  
 (g) any combination of (a) to (f).  
 
     
     
       3. A synthetic peptide according to  claim 2 , selected from the group consisting of:
 (pep1) Ile-Val-Leu  
 (pep4) Asn-Ile-Asn-Val-Ile-Val-Leu (SEQ ID NO:3),  
 (pep5) Ile-Val-Leu-Glu-Leu-Lys-Gly (SEQ ID NO:4),  
 (pep6) Asn-Val-Ile-Val-Leu (SEQ ID NO:5)  
 (pep7) Ala-Val-Leu  
 (pep8) Ile-Ala-Leu  
 (pep9) Ile-Val-Ala  
 (pep10) Glu-Val-Leu  
 (pep11, linear) and (pep12, cyclic) Cys-Ile-Val-Leu-Ala-Cys (SEQ ID NO:6) and,  
 (pep13, linear) and (pep14, cyclic) Cys-Ile-Val-Leu-Ala-Ala-Cys (SEQ ID NO:7).  
 
     
     
       4. The synthetic peptide Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:1) (pep2), and derivatives thereof according to  claim 1 , obtained by:
 (a) elongation at the C- and/or N-terminal ends by up to 4 further amino acid residues total;  
 (b) substitution of the Glu residue by a natural or non-natural charged or polar charged amino acid residue (c) substitution of the Phe residue by a natural or non-natural hydrophobic aliphatic or aromatic amino acid residue;  
 (d) substitution of the Leu residue by a natural or non-natural hydrophobic aliphatic or aromatic amino acid residue;  
 (e) substitution of the Asn residue by a hydrophilic, non-charged, aliphatic natural or non-natural amino acid residue;  
 (f) substitution of the Arg residue by a positively charged, natural or non-natural amino acid residue;  
 (g) substitution of the Trp residue by a natural or non-natural hydrophobic, aliphatic or aromatic, amino acid residue;  
 (h) substitution of the Ile residue by a natural or non-natural hydrophobic, aliphatic or aromatic, amino acid residue;  
 (i) substitution of the Thr residue by an aliphatic hydrophobic amino acid residue or a hydroxy- or thio-containing amino acid residue;  
 (j) truncation by up to 4 amino acid residues from either the C or N terminal;  
 (k) amidation of the C-terminal Thr;  
 (l) cyclization of pep2 or of any peptide of (a) to (k); or  
 (m) any combination of (a) to (l).  
 
     
     
       5. A peptide according to  claim 4 , selected from the group consisting of:
 (pep2) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:1)  
 (pep15) Ile-Val-Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:8)  
 (pep16) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr-Phe-Cys (SEQ ID NO:9)  
 (pep17) Ala-Thr-Ile-Val-Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:10)  
 (pep18) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr-Phe-Cys-Gln-Ser (SEQ ID NO:11)  
 (pep19) Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:12)  
 (pep20) Arg-Trp-Ile-Thr (SEQ ID NO:13)  
 (pep21) Glu-Phe-Leu-Asn (SEQ ID NO:14)  
 (pep22) Ala-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:15)  
 (pep23) Lys-Phe-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:16)  
 (pep24) Glu-Ala-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:17)  
 (pep25) Glu-Val-Leu-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:18)  
 (pep26) Glu-Phe-Ala-Asn-Arg-Trp-Ile-Thr (SEQ ID NO:19)  
 (pep27) Glu-Phe-Leu-Ala-Arg-Trp-Ile-Thr (SEQ ID NO:20)  
 (pep28) Glu-Phe-Leu-Asn-Ala-Trp-Ile-Thr (SEQ ID NO:21)  
 (pep29) Glu-Phe-Leu-Asn-Glu-Trp-Ile-Thr (SEQ ID NO:22)  
 (pep30) Glu-Phe-Leu-Asn-Arg-Ala-Ile-Thr (SEQ ID NO:23)  
 (pep31) Glu-Phe-Leu-Asn-Arg-Trp-Ala-Thr (SEQ ID NO:24)  
 (pep32) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Ala (SEQ ID NO:25)  
 (pep33) Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr-NH 2  (SEQ ID NO:26) and,  
 (pep34, linear) and (pep35, cyclic) Cys-Glu-Phe-Leu-Asn-Arg-Trp-Ile-Thr-Ala-Cys (SEQ ID NO:27).  
 
     
     
       6. The synthetic peptide Arg-Met-Leu-Thr (SEQ ID NO:2) (pep3), and derivatives thereof according to  claim 1 , obtained by:
 (a) elongation by up to 4 further amino acid residues at the C and/or N terminal end;  
 (b) substitution of the Arg residue by a natural or non-natural positively charged amino acid residue;  
 (c) substitution of the Met residue by a natural or non-natural hydrophobic, aliphatic or aromatic, amino acid residue;  
 (d) substitution of the Leu residue by a natural or non-natural hydrophobic, aliphatic or aromatic, amino acid residue;  
 (e) substitution of the Thr residue by an aliphatic hydrophobic amino acid residue or a hydroxy- or thio-containing amino acid residue;  
 (f) amidation of the C-terminal Thr residue;  
 (g) cyclization of pep3 or of any peptide of (a) to (f); or  
 (h) any combination of (a) to (g).  
 
     
     
       7. A peptide according to  claim 6 , selected from the group consisting of:
 (pep3) Arg-Met-Leu-Thr (SEQ ID NO:2)  
 (pep36) Ala-Met-Leu-Thr (SEQ ID NO:28)  
 (pep37) Arg-Ala-Leu-Thr (SEQ ID NO:29)  
 (pep38) Arg-Met-Ala-Thr (SEQ ID NO:30)  
 (pep39) Arg-Met-Leu-Ala (SEQ ID NO:31)  
 (pep40) Lys-Met-Leu-Thr (SEQ ID NO:32)  
 (pep41) Arg-Val-Leu-Thr (SEQ ID NO:33)  
 (pep42) Arg-Met-Leu-Thr-NH 2  (SEQ ID NO:34)  
 (pep43) Pro-Lys-Leu-Thr-Arg-Met-Leu-Thr (SEQ ID NO:35)  
 (pep44) Arg-Met-Leu-Thr-Phe-Lys-Phe-Tyr (SEQ ID NO:36) and,  
 (pep45, linear) and (pep46, cyclic) Cys-Arg-Met-Leu-Thr-Ala-Cys (SEQ ID NO:37).  
 
     
     
       8. A pharmaceutical composition comprising at least one synthetic peptide or peptide derivative according to  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
       9. A method for the treatment and/or alleviation of chronic inflammatory disorders comprising administering to a subject in need thereof an effective amount of an anti-inflammatory synthetic peptide according to  claim 1 . 
     
     
       10. The synthetic peptide of  claim 1 , which is pep2 (SEQ ID NO:1). 
     
     
       11. A pharmaceutical composition comprising the synthetic peptide of  claim 10  and a pharmaceutically acceptable carrier. 
     
     
       12. A method for the treatment and/or alleviation of chronic inflammatory disorders comprising administering to a subject in need thereof an effective amount of an anti-inflammatory synthetic peptide according to  claim 10 . 
     
     
       13. The synthetic peptide and derivatives thereof according to  claim 4 , wherein:
 said elongation is according to the natural sequence of IL-2;  
 said substitution of the Glu residue is selected from the group consisting of Lys, Arg, Asp, Gln, and Asn;  
 said substitution of the Phe residue is selected from the group consisting of Ala, Val, Ile, Leu, Tyr, Trp, Phe, Met, and Nle;  
 said substitution of the Leu residue is selected from the group consisting of Ala, Val, Ile, Leu, Tyr, Trp, Phe, Met, and Nle;  
 said substitution of the Asn residue is Gln;  
 said substitution of the Arg residue is selected from the group consisting of Lys, Orn, and homoArg;  
 said substitution of the Trp residue is selected from the group consisting of Tyr, Ile, Leu, Nle, Tic, Phe, 4-phenyl-Phe, and 4-methyl-Phe;  
 said substitution of the Ile residue is selected from the group consisting of Tyr, Phe, Leu, Nle, and Tic; and  
 said substitution of the Thr residue is selected from the group consisting of Ala, Ile, Leu, Cys, and Ser.  
 
     
     
       14. The synthetic peptide and derivatives thereof according to  claim 2 , wherein:
 said elongation is according to the natural sequence of IL-2;  
 said substitution of the Ile residue is selected from the group consisting of Glu, Asp, Asn, Gln, Ala, and Val;  
 said substitution of the Val residue is selected from the group consisting of Ala, Ile, Leu, Met, Nle, and Phe; and  
 said substitution of the Leu residue is selected from the group consisting of Ala, Ile, Met, Nle, Phe, and Val.  
 
     
     
       15. The synthetic peptide and derivatives thereof according to  claim 6 , wherein:
 said elongation is according to the natural sequence of IL-2;  
 said substitution of the Arg residue is selected from the group consisting of Lys, Orn, homoArg, and diaminobutyric acid;  
 said substitution of the Met residue is selected from the group consisting of Phe, Tyr, Ile, Leu, Nle, and Tic;  
 said substitution of the Leu residue is selected from the group consisting of Phe, Tyr, Nle, and Tic; and  
 said substitution of the Thr residue is selected from the group consisting of Ala, Ile, Leu, Ser, and Cys.